1. Field of Invention
The present invention relates a kit and a cleavage detection apparatus, and, in particular, a kit and a cleavage detection apparatus with high sequence specificity. The present invention also relates a gene therapy by using the nucleic acid cleavage kit.
2. Related Art
Nucleic acid cleavage is a reaction of breakage in the nucleic acid sequence imposed by external force. It can be applied to clinical healthcare, biotechnology or other related fields. Previously, various types of conventional nucleic acid cleavage tools have been used. Although they may be slightly different in composition, restriction enzymes with high specificity have been widely used in recognition and cleavage on nucleic acids.
However, as the expansion of the application scope of nucleic acid cleavage, the sequence-specificity provided by restriction enzymes has been insufficient to meet the demand. For example, several important researches and medical fields, such as gene transformation, genetic mapping and gene therapy, are fairly expecting the development of a highly specific nucleic acid cleavage tool.
In particular, the lengths of the nucleic acid sequences in higher organisms like human beings are approximately 105-107 base pairs. To create a site-specific cleavage on these nucleic acids, it has to use a cleavage tool possessing the ability of recognizing at least 8 to 15 base pairs. However, conventional restriction enzymes cannot recognize more than 8 base pairs and are incompetent for nucleic acid cleavage. Similarly, for implementing a genetic analysis on a genome (generally about 105-107 base pairs in length) or nucleic acids with high sequence similarity (for example heterochromatin), the basic demand of the sequence-specific recognition ability provided by a nucleic acid cleavage tool should be more than 8 base pairs correspondingly. The conventional nucleic acid cleavage tools used restriction enzymes as the functional components are certainly insufficient for certain application.
Besides, in order to improve the ability of nucleic acid sequence recognition, conventional methods have indicated oligonucleotides, which can bind to other nucleic acid segments containing corresponding sequences, can provide highly sequence-specific recognition for particular nucleic acid sequences. However, certain feature of the oligonucleotides was only applied as a nucleic acid probe for detecting whether particular sequences exist in an organism but not applied in cooperating with nucleic acid cleavage technique.
Therefore, it is an important subject of the invention to provide a nucleic acid cleavage tool with high sequence-specificity.